抗生素测定培养基L

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规格: 250g 500g

联系方式:I47-825O-882O

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莫能菌素测定培养基产品基本信息

产品名称: 抗生素测定培养基L;莫能菌素测定培养基
英文名称: Antibiotic Assay Medium L
培养基类型: 非选择性培养基
级别: for microbiology
品牌: ELITE-MEDIA(艾礼培养基)
产品目录号: M295-01、M295-02
产品规格: 250g、500g
产品外观: 麦秸色粉末。
灭菌后颜色与澄清度: 浅黄色略不透明凝胶。
保存条件: 密封,2-25°C保存。
注意事项: 避免摄入、吸入、皮肤接触。
相关产品: --
艾礼培养基


产品描述:

抗生素测定培养基L-AOAC(Antibiotic Assay Medium L, AOAC)作为底层琼脂,通过琼脂扩散法测定莫能菌素(Monensin)效价。测试微生物为枯草芽孢杆菌ATCC6633(Bacillus subtilis ATCC6633)。本配方是AOAC推荐配方。


配方与配制方法

成分 g/L
磷酸氢二钾 0.69
磷酸二氢钾 0.45
酵母提取物 2.5
无水葡萄糖 10
琼脂 15
终溶液pH 6.0± 0.2

配制方法:
1. 称取28.64g本产品,用1000ml纯水重悬。
2. 加热使培养基完全溶解。
3. 121°C 灭菌15min。
4. 冷却至45-50°C 时倒平板。


实验方法

Use single inoculated agar layer. Optimum concentration of suspension of Bacillus subtilis , is determined before assay by preparing trial plates. Usually 0.5 ml suspension is used per 100 ml of seed agar, to obtain appropriate inhibition zones (17.5 ± 2.5 mm with 0.5µg/ml). For actual assay add appropriate amount of suspension to sterile, molten medium, mix and pour 6 ml into sterile Petri plate. Cover and refrigerate for about 1 hour before use.

For the standard graph or response lines prepare dilution using 50% methanol to obtain 0.25, 0.5, 1.0 and 2.0 µg monensin/ml. Reference concentration is 0.5 µg/ml. To obtain standard curve 10 seeded agar plates are used placed with cylinders. Different standard concentrations are filled in it. Incubate at 16 18 hours at 35-37°C and measure diameters of zones of inhibition. Weigh 20gram finished feed and 5 gram premix and add it to chromatographic column. Elute with 9:1 methanol water. 200 ml elute is again diluted with 50% methanol to 0.5µg monensin /ml. This is called assay solution. Use 5 plates for each assay solution. Fill the alternate cylinders with reference concentration and assay solution after incubation at 35-37°C for 16-18 hours, measure diameters of zones of inhibition to nearest 0.1 mm. Average 10 reading of reference concentration and 10 reading of reference concentration and 10 readings of assay solution.

If assay solution gives larger average than reference concentration add difference between them to reference point on standard curve. If assay solution gives smaller value than reference concentration, substract difference between them from reference point. Using corrected value of assay solution determine amount of antibiotic.

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